Crystalline forms of modified triterpenoid hydrochloride salts with hiv maturation inhibitor activity

ABSTRACT

The present invention relates to crystalline forms of modified triterpenoid hydrochloride salts with HIV maturation inhibitor activity. More specifically, the present invention relates to the H-4, T1F-1, and T1F-2 crystalline forms. The present invention also relates to pharmaceutical compositions comprising these crystalline forms, as well as to methods of using these crystalline forms for the treatment of the HIV virus, and methods for obtaining these crystalline forms.

RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent Application Ser. No. 62/147,237 filed on Apr. 14, 2015, the disclosure of which is incorporated by reference herein in its entirety.

FIELD OF THE INVENTION

The present invention relates to crystalline forms of modified triterpenoid hydrochloride salts with HIV maturation inhibitor activity. More specifically, the present invention relates to the following three crystalline forms: form H-4, which is a crystalline monohydrochloride salt monohydrate, corresponding to the IUPAC name, H-4 form of 4-((1R,3aS,5aR,5bR, 7aR, 11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8, 11,11a,11b, 12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)benzoic acid hydrochloride hydrate, and to forms T1F-1 and T1F-2, which are both crystalline monohydrochloride salts, corresponding to the IUPAC name 4-((1R,3aS,5aR,5bR, 7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothio morpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b, 12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)benzoic acid hydrochloride. The present invention also relates to pharmaceutical compositions comprising these crystalline forms, as well as to methods of using these crystalline forms for the treatment of the HIV virus, and methods for obtaining these crystalline forms.

BACKGROUND OF THE INVENTION

HIV-1 (human immunodeficiency virus-1) infection and AIDS (acquired immune deficiency syndrome) remains a major medical problem. According to UNAIDS, at the end of 2014 nearly 37 million people were living with HIV. The number of cases of HIV has risen rapidly. In 2005, approximately 5.0 million new infections were reported, and 3.1 million people died from AIDS. Therefore, novel anti-HIV agents exhibiting distinct resistance patterns, and favorable pharmacokinetic as well as safety profiles are needed to provide more treatment options.

An emerging class of compounds for the treatment of HIV are called HIV maturation inhibitors. Maturation is the last of as many as ten or more steps in HIV replication or the HIV life cycle, in which HIV becomes infectious as a consequence of several HIV protease-mediated cleavage events in the gag protein that ultimately results in release of the capsid (CA) protein. Maturation inhibitors prevent the HIV capsid from properly assembling and maturing, from forming a protective outer coat, or from emerging from human cells. Instead, non-infectious viruses are produced, preventing subsequent cycles of HIV infection.

The HIV maturation triterpenoid compound of Formula I, below:

has been set forth and described in PCT Patent Application Publication No. WO 2012/106190 A1, published Aug. 9, 2012, and its US equivalents US Patent Application No. US 2013/0035318 A1, published Feb. 7, 2013 and U.S. Pat. No. 8,846,647 B2, issued Sep. 30, 2014, which are incorporated herein by reference. These documents describe and set forth various methods for making the compound of Formula I. This compound is also known by the IUPAC name 4-((1R,3 aS,5aR,5bR,7aR,11aS,11bR, 13 aR,13bR)-3a-((2-(1,1-dioxidothio morpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6, 7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl) benzoic acid.

What is now needed in the art are one or more new methods for reproducibly producing, stable crystalline forms of this triterpenoid HIV maturation inhibitor having desirable properties for treatment of HIV and incorporation into pharmaceutical compositions.

It has been found that certain forms of the hydrochloride salt of the compound represented by Formula I, and herein referred to as Compound I, can each be repeatedly prepared and crystallized into three particular forms, herein referred to as Form H-4 (which is a monohydrate form), Form T1F-1, and Form T1F-2.

These crystalline forms of Compound I offer excellent purification capacity and desirable formulation properties for incorporation into pharmaceutical compositions.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 illustrates experimental and simulated powdered X-Ray diffraction patterns (CuKα λ=1.54178 Å at T=room temperature, approximately 25° C.) of the H-4 crystalline form of Compound I. The data is presented as intensity versus 20 (two theta) in degrees. The experimental pattern is shown in the top part of the plot and the simulated in the bottom part of the plot.

FIG. 2 illustrates the differential scanning calorimetry pattern of the H-4 crystalline form of Compound I. The data is presented as heat flow in Watts (W) per gram versus temperature in ° C.

FIG. 3 illustrates the thermogravimetric analysis pattern of the H-4 crystalline form of Compound I. The data is presented as weight percent versus temperature in ° C.

FIG. 4 illustrates the Raman spectrum of the H-4 crystalline form of Compound I. The data is presented as Raman intensity versus Raman shift (cm⁻¹).

FIG. 5 illustrates the FTIR spectrum of the H-4 crystalline form of Compound I. The data is presented as log(1/R) versus wavenumbers (cm⁻¹).

FIG. 6 illustrates experimental and simulated powdered X-Ray diffraction patterns (CuKα λ=1.54178 Å at T=room temperature, approximately 25° C.) of the T1F-1 crystalline form of Compound I. The data is presented as intensity versus 2θ (two theta) in degrees. The experimental pattern is shown in the top part of the plot and the simulated in the bottom part of the plot.

FIG. 7 illustrates the differential scanning calorimetry pattern of the T1F-1 crystalline form of Compound I. The data is presented as heat flow in Watts (W) per gram versus temperature in ° C.

FIG. 8 illustrates the thermogravimetric analysis pattern of the T1F-1 crystalline form of Compound I. The data is presented as weight percent versus temperature in ° C.

FIG. 9 illustrates experimental and simulated powdered X-Ray diffraction patterns (CuKα λ=1.54178 Å at T=room temperature, approximately 25° C.) of the T1F-2 crystalline form of Compound I. The data is presented as intensity versus 2θ (two theta) in degrees. The experimental pattern is shown in the top part of the plot and the simulated in the bottom part of the plot.

FIG. 10 illustrates the differential scanning calorimetry pattern of the T1F-2 crystalline form of Compound I. The data is presented as heat flow in Watts (W) per gram versus temperature in ° C.

FIG. 11 illustrates the thermogravimetric analysis pattern of the T1F-2 crystalline form of Compound I. The data is presented as weight percent versus temperature in ° C.

FIG. 12 illustrates the Raman spectrum of the T1F-2 crystalline form of Compound I. The data is presented as Raman intensity versus Raman shift (cm⁻¹).

FIG. 13 illustrates the FTIR spectrum of the T1F-2 crystalline form of Compound I. The data is presented as log(1/R) versus wavenumbers (cm⁻¹).

SUMMARY OF THE INVENTION

The present invention relates to crystalline forms of modified triterpenoid hydrochloride salts with HIV maturation inhibitor activity. More specifically, the present invention relates to the H-4, T1F-1, and T1F-2 forms. The H-4 form is a monohydrochloride monohydrate crystalline salt form and can be designated by the IUPAC name: 4-((1R,3aS, 5aR,5bR,7aR,11aS,11bR, 13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a, 8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)benzoic acid hydrochloride hydrate.

The T1F-1 and T1F-2 polymorphic monohydrohloride salt crystalline forms can be designated by the same IUPAC name 4-((1R,3aS,5aR,5bR,7aR,11aS,11bR, 13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)benzoic acid hydrochloride. For the sake of convenience, the same general chemical formula is shown for the T1F-1 and T1F-2 forms, recognizing that they are distinct crystalline forms.

The present invention also relates to pharmaceutical compositions comprising these crystalline forms, as well as to methods of using these crystalline forms for the treatment of the HIV virus, and methods for obtaining these crystalline forms.

DETAILED DESCRIPTION OF THE INVENTION

In a first embodiment, the invention is directed to form H-4, T1F-1, or T1F-2 of Compound I.

In another embodiment, the invention is directed to form H-4 of Compound I

In another embodiment, the invention is directed to form H-4 of 4-((1R,3aS,5aR,5bR, 7aR, 11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothio morpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8, 11,11a,11b, 12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)benzoic acid hydrochloride hydrate, and specifically to the monohydrochloride monohydrate form.

In another embodiment, the invention is directed to form H-4 of Compound I characterized by the following unit cell parameters:

Cell Dimensions:

a=8.9877(3) Å

b=12.7723(4) Å

c=17.9546(5) Å

α=90 degrees

β=96.875(2) degrees

γ=90 degrees

Space group Monoclinic, P2₁

Molecules/unit cell 2,

wherein measurement of said crystalline form is at a temperature between about 20° C. to about 25° C.

In another embodiment, the invention is directed to form H-4 of Compound I of characterized by fractional atomic coordinates within the unit cell as listed in Table 4.

In another embodiment, the invention is directed to form H-4 of Compound I with characteristic peak positions in the powder X-Ray diffraction pattern at values of two theta of 5.0±0.2, 8.5±0.2, 12.6±0.2, 13.9±0.2, 14.9±0.2, 17.1±0.2, and 18.3±0.2 at a temperature between about 20° C. and about 25° C., and in further embodiments, the invention is directed to a form with at least one of these characteristic peak positions, with one or more of these characteristic peak positions, with two or more of these characteristic peak positions, with three or more of these characteristic peak positions, with four or more of these characteristic peak positions, with five or more of these characteristic peak positions, or with six or more of these characteristic peak positions.

In another embodiment, the invention is directed to form H-4 of Compound I characterized by an experimental powder X-ray diffraction pattern substantially in accordance with that shown in FIG. 1.

In another embodiment, the invention is directed to form H-4 of Compound I characterized by a differential scanning calorimetry thermogram substantially in accordance with that shown in FIG. 2.

In another embodiment, the invention is directed to Form H-4 of Compound I having a broad endotherm observed up to about 150° C. in the thermogram.

In another embodiment, the invention is directed to form H-4 of Compound I according characterized by a thermogravimetric analysis plot substantially in accordance with that shown in FIG. 3.

In another embodiment, the invention is directed to form H-4 of Compound I characterized by about a 2.4 percent weight loss on heating up to about 150° C.

In another embodiment, the invention is directed to form H-4 of Compound I characterized by a Raman spectrum substantially in accordance with that shown in FIG. 4.

In another embodiment, the invention is directed to form H-4 of Compound I characterized by a Raman spectrum with characteristic peaks as listed in Table 8, and in further embodiments, the invention is directed to a form with at least one of these characteristic peaks, with one or more of these characteristic peaks, with two or more of these characteristic peaks, with three or more of these characteristic peaks, with four or more of these characteristic peaks, or with five or more of these characteristic peaks.

In another embodiment, the invention is directed to form H-4 of Compound I characterized by a FTIR spectrum substantially in accordance with that shown in FIG. 5.

In another embodiment, the invention is directed to form H-4 of Compound I characterized by a FTIR spectrum with characteristic absorbance bands as listed in Table 10, and in further embodiments, the invention is directed to a form with at least one of these absorbance bands, with one or more of these absorbance bands, with two or more of these absorbance bands, with three or more of these absorbance bands, or with four or more of these absorbance bands.

In another embodiment, the invention is directed to a substantially pure form of H-4 of Compound I.

In another embodiment, the invention is directed to a substantially pure form of H-4 of Compound I having a purity of at least 95 weight percent.

In another embodiment, the invention is directed to a substantially pure form of H-4 of Compound I having a purity of at least 99 weight percent.

In another embodiment, the invention is directed to a pharmaceutical composition comprising form H-4 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of H-4 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of H-4 of Compound I having a purity of at least 95 weight percent and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of H-4 of Compound I having a purity of at least 99 weight percent and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a pharmaceutical composition comprising Form H-4 of Compound I in combination with one or two additional compounds having anti-HIV activity.

In another embodiment, the invention is directed to a method of treating HIV infection in a mammal comprising administering to the mammal a therapeutically-effective amount of Form H-4 of Compound I.

In another embodiment, the invention is directed to such a method wherein the mammal is a human.

In another embodiment, the invention is directed to a method of treating HIV infection in a mammal comprising administering to the mammal a therapeutically-effective amount of a pharmaceutical composition comprising form H-4 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to such a method wherein the mammal is a human.

In another embodiment, the invention is directed to a method for preparing Form H-4 of Compound I

comprising: (1) contacting compound 9

with aqueous base to effect hydrolysis of the methyl ester to yield an in situ hydrolysis product, and (2) treating the in situ hydrolysis product with hydrochloric acid in a solvent system comprising a mixture of tetrahydrofuran, water, and acetonitrile to yield form H-4 of Compound I.

In another embodiment, the invention is directed to form T1F-1 of Compound I

In another embodiment, the invention is directed to form T1F-1 of 4-((1R,3aS,5aR,5bR, 7aR, 11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothio morpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8, 11,11a,11b, 12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)benzoic acid hydrochloride, and specifically to the monohydrochloride form.

In another embodiment, the invention is directed to form T1F-1 of Compound I characterized by the following unit cell parameters:

Cell Dimensions:

a=17.3307(6) Å

b=30.7745(9) Å

c=7.8210(2) Å

α=90 degrees

β=90 degrees

γ=90 degrees

Space group Orthorhombic, P2₁2₁2

Molecules/unit cell 4,

wherein measurement of said crystalline form is at a temperature between about 20° C. to about 25° C.

In another embodiment, the invention is directed to form T1F-1 of Compound I characterized by fractional atomic coordinates within the unit cell as listed in Table 5.

In another embodiment, the invention is directed to form T1F-1 of Compound I with characteristic peak positions in the powder X-Ray diffraction pattern at values of two theta of 7.7±0.2, 11.7±0.2, 13.7±0.2, 14.2≥0.2, 15.3±0.2, 16.4±0.2, and 17.6±0.2 at a temperature between about 20° C. and about 25° C., and in further embodiments, the invention is directed to a form with at least one of these characteristic peak positions, with one or more of these characteristic peak positions, with two or more of these characteristic peak positions, with three or more of these characteristic peak positions, with four or more of these characteristic peak positions, with five or more of these characteristic peak positions, or with six or more of these characteristic peak positions.

In another embodiment, the invention is directed to form T1F-1 of Compound I characterized by an experimental powder X-ray diffraction pattern substantially in accordance with that shown in FIG. 6.

In another embodiment, the invention is directed to form T1F-1 of Compound I characterized by a differential scanning calorimetry thermogram substantially in accordance with that shown in FIG. 7.

In another embodiment, the invention is directed to form T1F-1 of Compound I having a broad endotherm near about 275° C. in the thermogram.

In another embodiment, the invention is directed to form T1F-1 of Compound I characterized by a thermogravimetric analysis plot substantially in accordance with that shown in FIG. 8.

In another embodiment, the invention is directed to form T1F-1 of Compound I characterized by a negligible, i.e, about zero percent, weight loss on heating up to about 150° C.

In another embodiment, the invention is directed to a substantially pure form of T1F-1 of Compound I.

In another embodiment, the invention is directed to a substantially pure form of T1F-1 of Compound I having a purity of at least 95 weight percent.

In another embodiment, the invention is directed to a substantially pure form of T1F-1 of Compound I having a purity of at least 99 weight percent.

In another embodiment, the invention is directed to a pharmaceutical composition comprising form T1F-1 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of T1F-1 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of T1F-1 of Compound I having a purity of at least 95 weight percent and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of T1F-1 of Compound I having a purity of at least 99 weight percent and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a pharmaceutical composition comprising Form T1F-1 of Compound I in combination with one or two additional compounds having anti-HIV activity.

In another embodiment, the invention is directed to a method of treating HIV infection in a mammal comprising administering to the mammal a therapeutically-effective amount of Form T1F-1 of Compound I according to claim 27.

In another embodiment, the invention is directed to such a method of wherein the mammal is a human.

In another embodiment, the invention is directed to a method of treating HIV infection in a mammal comprising administering to the mammal a therapeutically-effective amount of a pharmaceutical composition comprising form T1F-1 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to such a method wherein the mammal is a human.

In another embodiment, the invention is directed to a method for preparing Form T1F-1 of Compound I

comprising: (1) contacting compound 9

with aqueous base to effect hydrolysis of the methyl ester to yield an in situ hydrolysis product, and (2) treating the in situ hydrolysis product with hydrochloric acid in a solvent system comprising a mixture of ethanol and water to yield form T1F-1 of Compound I.

In another embodiment, the invention is directed to form T1F-2 of Compound I

In another embodiment, the invention is directed to form T1F-2 of 4-((1R,3aS,5aR,5bR, 7aR, 11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothio morpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8, 11,11a,11b, 12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)benzoic acid hydrochloride, and specifically to the monohydrochloride form.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by the following unit cell parameters:

Cell Dimensions:

a=43.967(1) Å

b=7.7218(3) Å

c=12.0227(4) Å

α=90 degrees

β=90.045(2) degrees

γ=90 degrees

Space group Monoclinic, C2

Molecules/unit cell 4,

wherein measurement of said crystalline form is at a temperature between about 20° C. to about 25° C.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by fractional atomic coordinates within the unit cell as listed in Table 6.

In another embodiment, the invention is directed to form T1F-2 of Compound I with characteristic peaks in the powder X-Ray diffraction pattern at values of two theta of 4.1±0.2, 7.4±0.2, 11.9±0.2, 13.0±0.2, 14.0±0.2, 14.5±0.2, and 15.5±0.2 at a temperature between about 20° C. and about 25° C., and in further embodiments, the invention is directed to a form with at least one of these characteristic peak positions, with one or more of these characteristic peak positions, with two or more of these characteristic peak positions, with three or more of these characteristic peak positions, with four or more of these characteristic peak positions, with five or more of these characteristic peak positions, or with six or more of these characteristic peak positions.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by an experimental powder X-ray diffraction pattern substantially in accordance with that shown in FIG. 9.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by a differential scanning calorimetry thermogram substantially in accordance with that shown in FIG. 10.

In another embodiment, the invention is directed to form T1F-2 of Compound I having a broad endotherm near about 275° C. in the thermogram.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by a thermogravimetric analysis plot substantially in accordance with that shown in FIG. 11.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by a negligible, i.e, about zero percent, weight loss on heating up to about 150° C.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by a Raman spectrum substantially in accordance with that shown in FIG. 12.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by a Raman spectrum with characteristic peaks as listed in Table 9. and in further embodiments, the invention is directed to a form with at least one of these characteristic peaks, with one or more of these characteristic peaks, with two or more of these characteristic peaks, with three or more of these characteristic peaks, with four or more of these characteristic peaks, or with five or more of these characteristic peaks.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by a FTIR spectrum substantially in accordance with that shown in FIG. 13.

In another embodiment, the invention is directed to form T1F-2 of Compound I characterized by a FTIR spectrum with characteristic absorbance bands as listed in Table 11, and in further embodiments, the invention is directed to a form with at least one of these absorbance bands, with one or more of these absorbance bands, with two or more of these absorbance bands, with three or more of these absorbance bands, or with four or more of these absorbance bands.

In another embodiment, the invention is directed to a substantially pure form of T1F-2 of Compound I.

In another embodiment, the invention is directed to a substantially pure form of T1F-2 of Compound I having a purity of at least 95 weight percent.

In another embodiment, the invention is directed to a substantially pure form of T1F-2 of Compound I having a purity of at least 99 weight percent.

In another embodiment, the invention is directed to a pharmaceutical composition comprising form T1F-2 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of T1F-2 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of T1F-2 of Compound I having a purity of at least 95 weight percent and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a substantially pure form of T1F-2 of Compound I having a purity of at least 99 weight percent and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to a pharmaceutical composition comprising Form T1F-2 of Compound I in combination with one or two additional compounds having anti-HIV activity.

In another embodiment, the invention is directed to a method of treating HIV infection in a mammal comprising administering to the mammal a therapeutically-effective amount of Form T1F-2 of Compound I.

In another embodiment, the invention is directed to such a method wherein the mammal is a human.

In another embodiment, the invention is directed to a method of treating HIV infection in a mammal comprising administering to the mammal a therapeutically-effective amount of a pharmaceutical composition comprising form T1F-2 of Compound I and a pharmaceutically acceptable carrier or diluent.

In another embodiment, the invention is directed to such a method wherein the mammal is a human.

In another embodiment, the invention is directed to a method for preparing Form T1F-2 of Compound I

comprising: (1) contacting compound 9

with aqueous base to effect hydrolysis of the methyl ester to yield an in situ hydrolysis product, and (2) treating the in situ hydrolysis product with hydrochloric acid in a solvent system comprising a mixture of isopropanol and water to yield form T1F-2 of Compound I.

Other embodiments of the present disclosure can comprise suitable combinations of two or more embodiments and/or aspects disclosed herein.

Yet other embodiments and aspects of the disclosure will be apparent according to the description provided below.

The compounds of the present disclosure also exist as tautomers; therefore the present disclosure also encompasses all tautomeric forms.

Definitions

As used herein “polymorph(s)” or “polymorphic form(s)” refers to crystalline forms having the same chemical composition but different spatial arrangements of the molecules, atoms, and/or ions forming the crystal.

The term “pharmaceutically acceptable,” as used herein, refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem complications commensurate with a reasonable benefit/risk ratio.

The term “substantially pure,” as used herein refers to a crystalline form of Compound I which is great than about 90% pure. This means that the particular crystalline form of Compound I does not contain more than about 10% of any other compound, and, in particular, does not contain more than about 10% of any other form of Compound I.

The term “therapeutically effective amount,” as used herein, is intended to include an amount of the crystalline forms of Compound I that is effective when administered alone or in combination to treat HIV.

The term “treating” refers to: (i) preventing a disease, disorder or condition from occurring in a patient which may be predisposed to the disease, disorder and/or condition but has not yet been diagnosed as having it; (ii) inhibiting the disease, disorder or condition, i.e., arresting its development; and/or (iii) relieving the disease, disorder or condition, i.e., causing regression of the disease, disorder and/or condition.

In various embodiments the disclosure provides crystalline forms of Compound I. These crystalline forms are the H-4 monhydrochloride monohydrate form and the T1F-1 and the T1F-2 monohydrochloride forms. The T1F-1 and the T1F-2 forms are non-solvated polymorphic forms of the monohydrochloride salt. These crystalline forms of Compound I can be employed in pharmaceutical compositions which can optionally include one or more other components selected, for example, from the group consisting of excipients, carriers, and one of other active pharmaceutical ingredients active chemical entities of different molecular structure.

In various embodiments the selected crystalline forms have phase homogeneity indicated by less than 10 percent, in other embodiments the crystalline forms have phase homogeneity indicated by less than 5 percent, and in other embodiments the crystalline forms have phase homogeneity indicated by less than 2 percent of the total peak area in the experimentally measured powder x-ray diffraction (PXRD) pattern arising from the extra peaks that are absent from the simulated PXRD pattern. In other embodiments the crystalline forms have phase homogeneity with less than 1 percent of the total peak area in the experimentally measured PXRD pattern arising from the extra peaks that are absent from the simulated PXRD pattern.

In other embodiments, compositions are provided consisting essentially of the desired crystalline form of Compound I. The composition of these embodiments can comprise at least 90 weight percent of the desired crystalline form of Compound I, based on the weight of Compound I in the composition. The remaining material comprises other form(s) of the compound and/or reaction impurities and/or processing impurities arising from the preparation of the desired crystalline form. The presence of reaction impurities and/or processing impurities can be determined by analytical techniques known in the art, such as, for example, chromatography, nuclear magnetic resonance spectroscopy, mass spectrometry, or infrared spectroscopy.

General Preparation of Crystalline Materials:

Crystalline forms can be prepared by a variety of methods, including for example, crystallization or recrystallization from a suitable solvent, sublimation, growth from a melt, solid state transformation from another phase, crystallization from a supercritical fluid, and jet spraying. Techniques for crystallization or recrystallization of crystalline forms from a solvent mixture include, for example, evaporation of the solvent, decreasing the temperature of the solvent mixture, crystal seeding a supersaturated solvent mixture of the molecule and/or salt, freeze drying the solvent mixture, and addition of antisolvents (countersolvents) to the solvent mixture. High throughput crystallization techniques can be employed to prepare crystalline forms including polymorphs. Crystals of drugs, including polymorphs, methods of preparation, and characterization of drug crystals are discussed in Solid-State Chemistry of Drugs, S. R. Byrn, R. R. Pfeiffer, and J. G. Stowell, 2^(nd) Edition, SSCI, West Lafayette, Ind. (1999).

For crystallization techniques that employ solvent, the choice of solvent or solvents is typically dependent upon one or more factors, such as solubility of the compound, crystallization technique, and vapor pressure of the solvent. Combinations of solvents can be employed, for example, the compound can be solubilized into a first solvent to afford a solution, followed by the addition of an antisolvent to decrease the solubility of the compound in the solution and to afford the formation of crystals. An antisolvent is a solvent in which the compound has low solubility.

In some methods to prepare crystals, a compound is suspended and/or stirred in a suitable solvent to afford a slurry, which can be heated to promote dissolution. The term “slurry”, as used herein, means a saturated solution of the compound, which can also contain an additional amount of the compound to afford a heterogeneous mixture of the compound and a solvent at a given temperature.

Seed crystals can be added to a crystallization mixture to promote crystallization. Seeding can be employed to control growth of a particular crystalline form or to control the particle size distribution of the crystalline product. Accordingly, calculation of the amount of seeds needed depends on the size of the seed available and the desired size of an average product particle as described, for example, in “Programmed Cooling of Batch Crystallizers,” J. W. Mullin and J. Nyvlt, Chemical Engineering Science, 1971, 26, 369-377. In general, seeds of small size are needed to control effectively the growth of crystals in the batch. Seeds of small size can be generated by sieving, milling, or micronizing of large crystals, or by micro-crystallization of solutions. Care should be taken that milling or micronizing of crystals does not result in any change in crystallinity of the desired crystal form (i.e., a change to an amorphous form or to another crystal or polymorph form).

A cooled crystallization mixture can be filtered under vacuum, and the isolated solids can be washed with a suitable solvent, such as cold recrystallization solvent, and dried under a nitrogen purge to afford the desired crystalline form. The isolated solids can be analyzed by a suitable spectroscopic or analytical technique, such as solid state nuclear magnetic resonance, differential scanning calorimetry, X-Ray powder diffraction, or the like, to assure formation of the preferred crystalline form of the product. The resulting crystalline form is typically produced in an amount of greater than about 70 weight percent isolated yield, preferably greater than 90 weight percent isolated yield, based on the weight of the compound originally employed in the crystallization procedure. The product can be co-milled or passed through a mesh screen to remove lumps from the product, if necessary.

Crystalline forms can be prepared directly from the reaction medium of the process for preparing Compound I. These can be achieved, for example, by employing in the final process step a solvent or a mixture of solvents from which Compound I can be crystallized. Alternatively, crystalline forms can be obtained by distillation or solvent addition techniques. Suitable solvents for this purpose include, for example, the aforementioned non-polar solvents and polar solvents, including protic polar solvents such as alcohols, and aprotic polar solvents such as ketones. In the case of the T1F-1 and T1F-2 forms, their preparation would typically involve a drying step. In such instances the drying step can be conducted at 60° C. or higher.

The presence of more than one crystalline form in a sample can be determined by techniques such as PXRD or solid state nuclear magnetic resonance spectroscopy (SSNMR). For example, the presence of extra peaks in an experimentally measured PXRD pattern when compared with a simulated PXRD pattern can indicate more than one crystalline form in the sample. The simulated PXRD can be calculated from single crystal X-Ray data. See Smith, D. K., “A FORTRAN Program for Calculating X-Ray Powder Diffraction Patterns,” Lawrence Radiation Laboratory, Livermore, Calif., UCRL-7196 (April 1963).

Relationship of the Forms:

Herein we describe three crystalline forms of the monohydrochloride salt, Compound I, that have been prepared and identified: the monohydrochloride monohydrate H-4 form and two monohydrochloride forms designated as T1F-1 and T1F-2. It is found that the T1F-2 form is thermodynamically more stable than T1F-1 form at ambient conditions. The critical water activity between the T1F-2 and H-4 forms is in the range 0.05-0.24.

All three of these forms can be accessed through form conversion of a monohydrochloride salt form of Compound I which can be obtained from the hydrolysis and acidification of an ester of the free base form of Compound I, such as the corresponding methyl ester free base, shown below, which is also designated as Compound 9.

Although the monohyhdrochloride salt of Compound I can be isolated and then used to prepare the desired H-4, T1F-1, and T1F-2 forms, these forms are more typically prepared from the in situ generated compound (i.e. the carboxylic acid) resulting from the hydrolysis of the methyl ester of the free base form of Compound I (Compound 9). This hydrolysis product, as thus generated in situ, is then directly converted to the desired H-4, T1F-1, or T1F-2 form of Compound I. Scheme A shows the relationship of Compound I to the H-4, T1F-1, and T1F-2 crystalline forms. Other forms are also shown in Scheme A. These include: P44, which is an anhydrous form obtained upon drying of form H-4. Form IPA-2, which is an isopropyl solvate of Compound I. that upon drying yields form TF1-2. A further solvate obtained from a THF/water or THF/water/acetonitrile solvent from Compound I, that upon during yields form TF1-2. A further solvate obtained from an ethanol/water solvent, that upon drying yields form TF1-1.

It is found that other appropriate solvents and solvent ratios, and conditions, e.g., such as temperature, can be used in the interconversion or preparation of the desired form.

Scheme A showing the relationship of Compound I to the H-4, TF1-1, TF1-2, and other forms.

As seen from Scheme A, the H-4 monohydrochloride monohydrate salt can be generated from the monohydrochloride salt form of Compound I using one of the following binary solvent mixtures: isopropyl alcohol (IPA)/water, acetone/water, acetonitrile (MeCN)/water, ethanol (EtOH)/water, or tetrahydrofuran(THF)/water. Additionally, the ternary solvent mixture THF/water/MeCN can be used. If the resultant H-4 form is dried at a temperature greater than 60° C., then it converts to an anhydrous form of H-4, designated as P44. However, it has been found that the H-4 form is sensitive to relative humidity, and can be dehydrated to the P44 form at a temperature as low as 30° C. under a stream of dry nitrogen gas. In practice, to minimize conversion to the P44 form, the drying can be conducted under a stream of humidified nitrogen gas.

The T1F-1 monohydrochloride salt form of Compound I can be made by slurrying the crude monohydrochloride salt Compound I in a mixture of ethanol (EtOH)/water to generate a solvated form, that is dried to yield the T1F-1 form. Alternatively, the T1F-1 form can be crystallized from the in situ generated form of Compound I from an ethanol/water mixture.

The T1F-2 monohydrochloride salt form of Compound I can be made by slurrying the crude monohydrocyloride salt of Compound I in a mixture of isopropyl alcohol (IPA)/water to generate an isopropyl alcohol (IPA) solvate, designated as IPA-2, and drying this solvate to yield the T1F-2 form. Alternatively, the T1F-2 monohydrochloride can be generated by slurrying the crude monohydrochloride salt of Compound I in s mixture of tetrahydrofuran (THF)/water or THF/water/acetonitrile (MeCN), to generate a solvated form, that is dried to yield the T1F-2 form.

Preparation of the Methyl Ester Free Base Form of Compound I (Also Known as Compound 9) and Hydrolysis and Acidification to Yield the Carboxylic Acid Compound I

As discussed above, the methyl ester free base form of Compound I (Compound 9) can be hydrolyzed and acidified, and depending on the process conditions, the resultant in-situ generated carboxylic acid and Compound I (resulting from acidification of the carboxylic acid) can be used to prepare the desired crystalline form such as H-4, T1F-1, or T1F-2.

The process for making Compound I, which can have a water of hydration associated with it depending on the conditions, can be described by the following steps:

which comprises: (1) oxidizing the starting compound

to yield compound

and (2) contacting the compound 1 with reagent 2a

in solution to yield the compound 2b

and (3a) contacting the compound 2b with compound 3a

along with PdCl2Xantphos and aq. K₃PO₄ in solvent to yield 3b

and (3b) contacting the compound 3b with hydroxylamine to produce the compound 4

and (4a) contacting the compound 4 with an oxidant to initially produce the compound

(4b) reaction of compound 5 with TFAA to produce compound 6b

and (4c) contacting the compound 6 with a base and heat to produce the compound 7 as a result of a skeletal rearrangement

and (5) contacting the compound 7 with compound 8

and base in solvent to yield compound 9 (the methyl ester free base form of Compound I)

and (6) contacting compound 9 with aqueous base to effect hydrolysis of the methyl ester to yield the free base from of the Compound of Formula I, which can be converted to the hydrochloride salt by treatment with HCl. It is noted that the hydrolysis product and this Compound I are generated in situ and then carried on, without isolation, to the H-4, T1F-1, or T1F-2 form of Compound I.

Conceptually, the process of the foregoing steps can also be summarized and illustrated according to the following exemplified, non-limiting Scheme B:

The following describes the steps outlined in Scheme B, above.

Step 1: Preparation of Compound 1

This process involves the oxidation of both hydroxyl groups present in the starting material (betulin) to produce the desired keto-aldehyde 1. Three oxidative processes have been developed to achieve the desired initial transformation: [1] A Moffatt Oxidation employing 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and dimethyl sulfoxide (DMSO) in dichloromethane as solvent and a catalytic acid. Various cabodiimides can be employed, including, for example, diisopropylcarbodiimide (DIC) and dicyclohexylcarbodiimide (DCC). The identity of the activating catalytic acid can also vary, and include p-toluene sulfonic acid (p-TSA), Pyridinium p-toluenesulfonate (PPTS), dichloroacetic acid, or H₃PO₄. Of these, PPTS is preferred. [2] An aerobic oxidation employing a copper (I)/dimethoxybipyridine/8-hydroxy-8-azabicyclo[3.2.1]octan-3-one/TEMPO/NMI catalyst system under O₂ or air in CH₃CN and DCM. Other ligands such as bipyridine or N-benzylimidazole can also be employed. Other possible catalysts include ABNO, keto-ABNO, AZADO, and AZADOL but 8-hydroxy-8-azabicyclo[3.2.1]octan-3-one is preferred. It should be noted, that the use of 8-hydroxy-8-azabicyclo[3.2.1]octan-3-one as the oxidation catalyst precursor is preferred over the nitrosyl radical precursors such as ABNO, keto-ABNO AZADO, or AZADOL for commercial manufacturing because of its ease of preparation (2 steps) and its improved stability profile. [3] A modified Parikh-Doering oxidation that uses SO₃.trimethylamine and DMSO in dichloromethane or THF as solvents in their presence of base. Although SO₃.trimethylamine is preferred in this transformation, various other activators can be employed such as SO₃.pyridine, SO₃-Et₃N and P₂O₅. In addition, other bases such as triethylamine (TEA), tetramethyl guanidine (TMG), diisopropylethylamine (DIPEA) and triisobutylamine can also be utilized, although 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) is preferred.

The reaction scheme can be summarized as follows:

Step 2: Preparation of Compound 2

The subsequent transformation comprises the conversion of keto-aldehyde compound 1 to the enol-triflate compound 2. In the optimized process, 1 and 2a are dissolved into a solvent (typically, tetrahydrofuran (THF) or methyl tert-butyl ether (MTBE)), cooled to about −5° C. The ketone in this mixture is then selectively enolized by the addition of a strong base. It is preferred to perform this reaction using the bases lithium diisopropyl amide (LDA) or M-HMDS (hexamethyldisilazane), where M=Na, K, or Li. Without being bound by any particular theory, there are two particular aspects of this reaction: (1) preference for one carbonyl group over the other (i.e. reaction with the ketone in the presence of the aldehyde), which eliminates the potential protecting group requirement, and (2) the utilization of the reagent 2a which enables a selective and non-cryogenic reaction to be conducted. Other triflating reagents such as triflic anhydride are preferably not utilized as they will react with the base used for enolization.

Step 3: Preparation of Compound 3

In this sub-step, boronic acid 3a is coupled to enol-triflate compound 2b via a palladium catalyzed Suzuki coupling reaction. The product intermediate shown below is telescoped through a sub step in which hydroxyl amine is condensed with the aldehyde moiety to give oxime 4 after crystallization from aqueous isopropyl alcohol (IPA). The preferred pH for this transformation is 7.0-7.5 and is obtained by a pH adjustment using a mild organic acid, such as acetic acid. This palladium mediated cross coupling preferably occurs under the influence of the Xantphos ligand, aq. K₃PO₄ as base, and THF as solvent.

The reaction scheme for step 3 can be summarized as follows:

In a further embodiment, compounds 3b and 4 can be represented more generically as:

wherein R¹ is selected from the group of -alkyl, -aryl, and substituted aryl, with methyl being preferred.

In a further embodiment, compound 3a can be represented more generically as:

wherein R¹ is selected from the group of -alkyl, -aryl, and substituted aryl, with methyl being preferred, and also wherein R₂ is either a boronic acid, pinacolboronate ester, or isopropyl boronate ester.

Step 4: Preparation of Compound 6

This is a three sub-step process involving: [1] the selective oxidation of the oxime (4) with a hypervalent iodine (III) reagent to generate the nitrile oxide (5), [2] selective hydration of the nitrile oxide using trifluoroacetic anhydride (TFAA, 0° C.) to produce bis-TFA-derivative 6a, [3] hydrolytic removal of the two TFA groups to give hydroxamic acid 6b. This selective sequence enables site specific oxidation (PIFA) and hydration (TFAA) of the oxime moiety to produce hydroxamic acid 6b in excellent yields (typically about 85-95% yield).

In this first sub step (Step 4a), [Bis(Trifluoroacetoxy)iodo]benzene (PIFA) is used preferably as the stoichiometric oxidant (about 1.0-1.25 eq) and is conducted at 0° C. to ambient temperature in acetonitrile, THF, acetone, or other ketone solvent. Acetone or THF are the preferred solvents. Other oxidants can be employed for the transformation of 4 to 5. These reagents can include: phenyliodine diacetate (PIDA), N-bromosuccinimide (NBS), N-chlorosuccinimide (NCS), Chloramine-T, I₂O₅, but PIFA is preferred. Step 4b is initiated by the addition of TFAA. The reaction is conducted between 0° C. and ambient temperature and in the same solvents as step 4a. After reaction completion, H₂O is added to quench the reaction as well as to facilitate the crystallization of hydroxamic 6b.

The reaction scheme for step 4 can be summarized as follows:

Step 5: Preparation of Compound 7

Step 5 comprises the base mediated decarboxylative rearrangement (Lossen rearrangement) of the hydroxamic acid (6b) to yield the primary amine intermediate 7. This base-mediated Lossen rearrangement (DBU, CH₃CN, 70° C.) is carried out under very mild conditions (DBU, CH₃CN, 70° C.). Applicants have surprisingly discovered that CH₃CN performs as both solvent and as an initiator for the desired rearrangement through a self-propagating mechanism.

Step 5 is catalyzed by the addition of a base. Various organic or inorganic bases can be used, but 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) is preferred. The reaction can be conducted in various solvents and co-solvents with acetonitrile being preferred, and at elevated temperatures (typically >70° C.). Typical yields range from about 90-95% overall yield. It is believed that this sequence (steps 4+5) represents a unique and unprecedented method for the introduction of a C-17 nitrogen on the biologically important betulin framework.

The reaction scheme for step 5 can be summarized as follows:

Step 6: Preparation of Compound 9 (Methyl Ester Free Base Form of Compound I)

This is the penultimate step comprising the N-alkylation of 7 with 8 using i-Pr₂NEt as base. The product 9, the methyl ester free base form, can then either be isolated or converted to various salt forms.

In a further embodiment of the invention, compound 8 can be represented more generically as:

wherein X can be any number of leaving groups, with OMs being preferred.

Step 7: Preparation of Compound I

This is the final step comprising the saponification of the methyl ester free base using aq. n-Bu₄NOH in IPA/H₂O or THF. Other bases including NaOH, KOH, and LiOH can be implemented in this process, but n-Bu₄NOH is preferred. Acidification with HCl provides that hydrochloride salt, Compound I, which can either be isolated or left in situ and converted to a specific crystalline form.

The desired crystalline from of Compound I, i.e. H-4, T1F-1, or T1F-2, is obtained depending on the choice of solvent mixture, conditions, and as applicable, the drying step. Some representative conditions are shown below for Step 7 in Scheme 7.

Characterization:

The forms of Compound I described herein can be characterized using various techniques, the operation of which are well known to those of ordinary skill in the art. Examples of characterization methods include, but are not limited to, single crystal X-Ray diffraction (XRD), powder X-Ray diffraction (PXRD), simulated powder X-Ray patterns (Yin, S.; Scaringe, R. P.; DiMarco, J.; Galella, M. and Gougoutas, J. Z., American Pharmaceutical Review, 2003, 6, 2, 80), differential scanning calorimetry (DSC), solid-state ¹³C NMR (Earl, W. L. and Van der Hart, D. L., J. Magn. Reson., 1982, 48, 35-54), Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), moisture sorption isotherms, thermal gravimetric analysis (TGA), differential scanning calorimetry (DSC), and hot stage techniques.

The forms can be characterized and distinguished using single crystal X-Ray diffraction, which is based on unit cell measurements of a single crystal of form N-2. A detailed description of unit cells is provided in Stout & Jensen, X-Ray Structure Determination: A Practical Guide, Macmillan Co., New York (1968), Chapter 3, which is herein incorporated by reference. Alternatively, the unique arrangement of atoms in spatial relation within the crystalline lattice can be characterized according to the observed fractional atomic coordinates. Another means of characterizing the crystalline structure is by powder X-Ray diffraction analysis in which the diffraction profile is compared to a simulated profile representing pure powder material, both run at the same analytical temperature, and measurements for the subject form characterized as a series of 2θ (two theta) values.

One of ordinary skill in the art will appreciate that an X-Ray diffraction pattern can be obtained with a measurement of error that is dependent upon the measurement conditions employed. In particular, it is generally known that intensities in an X-Ray diffraction pattern can fluctuate depending upon measurement conditions employed. It should be further understood that relative intensities can also vary depending upon experimental conditions, and, accordingly, the exact order of intensity should not be taken into account. Additionally, a measurement error of diffraction angle for a conventional X-Ray diffraction pattern is typically about 5 percent or less, and such degree of measurement error should be taken into account as pertaining to the aforementioned diffraction angles. Consequently, it is to be understood that the crystal forms of the present disclosure are not limited to the crystal forms that provide X-Ray diffraction patterns completely identical to the X-Ray diffraction patterns depicted in the accompanying Figures disclosed herein. Any crystal form that provides the X-Ray diffraction pattern, DSC thermogram, thermogravimetric analysis, Raman spectrum, or FTIR spectrum substantially identical to those disclosed in the accompanying Figures fall within the scope of the present disclosure. The ability to ascertain substantial identities of X-Ray diffraction patters is within the purview of one of ordinary skill in the art.

Utility:

The H-4, T1F-1, T1F-2, forms of Compound I can be used to treat or prevent HIV infection.

The active ingredient, i.e., the H-4, T1F-1, or T1F-2 form of Compound I, in such compositions typically comprises from 0.1 weight percent to 99.9 percent by weight of the composition, and often comprises from about 5 to 95 weight percent. In some cases, the pH of the formulation can be adjusted with pharmaceutically acceptable modifiers (such as calcium carbonate and magnesium oxide) to enhance the stability of the formulated compound or its delivery form. Formulations of the crystalline forms of the present disclosure can also contain additives for enhancement of absorption and bioavailability.

The compounds of the present invention, according to all the various embodiments described above, can be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques), by inhalation spray, or rectally, and by other means, in dosage unit formulations containing non-toxic pharmaceutically acceptable carriers, excipients and diluents available to the skilled artisan. One or more adjuvants can also be included.

Other suitable carriers for the above noted compositions can be found in standard pharmaceutical texts, e.g. in “Remington's Pharmaceutical Sciences”, 19th ed., Mack Publishing Company, Easton, Pa., 1995. Further details concerning the design and preparation of suitable delivery forms of the pharmaceutical compositions of the disclosure are known to those skilled in the art.

When the desired polymorph is formulated together with a pharmaceutically acceptable carrier, the resulting composition can be administered in vivo to mammals, such as man, to inhibit or to treat or prevent HIV virus infection.

Thus, in accordance with the present invention, there is further provided a method of treatment, and a pharmaceutical composition, for treating viral infections such as HIV infection and AIDS. The treatment involves administering to a patient in need of such treatment a pharmaceutical composition which contains an antiviral effective amount of the desired crystalline form of Compound I, together with one or more pharmaceutically acceptable carriers, excipients or diluents. As used herein, the term “antiviral effective amount” means the total amount of each active component of the composition and method that is sufficient to show a meaningful patient benefit, i.e., inhibiting, ameliorating, or healing of acute conditions characterized by inhibition of the HIV infection. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously. The terms “treat, treating, treatment” as used herein and in the claims means preventing, ameliorating or healing diseases associated with HIV infection.

The pharmaceutical compositions of the invention can be in the form of orally administrable suspensions or tablets; as well as nasal sprays, sterile injectable preparations, for example, as sterile injectable aqueous or oleaginous suspensions or suppositories. Pharmaceutically acceptable carriers, excipients or diluents can be utilized in the pharmaceutical compositions, and are those utilized in the art of pharmaceutical preparations.

When administered orally as a suspension, these compositions are prepared according to techniques typically known in the art of pharmaceutical formulation and can contain microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners/flavoring agents known in the art. As immediate release tablets, these compositions can contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrants, diluents, and lubricants known in the art.

When orally administered, the pharmaceutical compositions of this disclosure can be administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, and aqueous suspensions and solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, can also be added. For oral administration in a capsule form, useful carriers/diluents include lactose, high and low molecular weight polyethylene glycol, and dried corn starch. When aqueous suspensions are administered orally, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents can be added.

The compounds herein set forth can be administered orally to humans in a dosage range of about 1 to 100 mg/kg body weight in divided doses, usually over an extended period, such as days, weeks, months, or even years. One preferred dosage range is about 1 to 10 mg/kg body weight orally in divided doses. Another preferred dosage range is about 1 to 20 mg/kg body weight in divided doses. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient can be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

The pharmaceutical compositions can be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The details concerning the preparation of such compounds are known to those skilled in the art.

The injectable solutions or suspensions can be formulated according to known art, using suitable non-toxic, parenterally acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.

Also contemplated herein is the desired crystalline form of Compound I in combination with one or more other agents useful in the treatment of AIDS. For example, the compounds of this disclosure can be effectively administered, whether at periods of pre-exposure and/or post-exposure, in combination with effective amounts of the AIDS antivirals, immunomodulators, antiinfectives, or vaccines, such as those in the following non-limiting table:

Antivirals

Drug Name Manufacturer Indication 097 Hoechst/Bayer HIV infection, AIDS, AIDS related complex (ARC) (non-nucleoside reverse tran- scriptase (RT) inhibitor) Amprenavir Glaxo Wellcome HIV infection, 141 W94 AIDS, ARC GW 141 (protease inhibitor) Abacavir (1592U89) Glaxo Wellcome HIV infection, GW 1592 AIDS, ARC (RT inhibitor) Acemannan Carrington Labs ARC (Irving, TX) Acyclovir Burroughs Wellcome HIV infection, AIDS, ARC AD-439 Tanox Biosystems HIV infection, AIDS, ARC AD-519 Tanox Biosystems HIV infection, AIDS, ARC Adefovir dipivoxil Gilead Sciences HIV infection AL-721 Ethigen ARC, PGL (Los Angeles, CA) HIV positive, AIDS Alpha Interferon Glaxo Wellcome Kaposi's sarcoma, HIV in combination w/Retrovir Ansamycin Adria Laboratories ARC LM427 (Dublin, OH) Erbamont (Stamford, CT) Antibody which Advanced Biotherapy AIDS, ARC Neutralizes pH Concepts Labile alpha aberrant (Rockville, MD) Interferon AR177 Aronex Pharm HIV infection, AIDS, ARC Beta-fluoro-ddA Nat'l Cancer Institute AIDS-associated diseases BMS-234475 Bristol-Myers Squibb/ HIV infection, (CGP-61755) Novartis AIDS, ARC (protease inhibitor) CI-1012 Warner-Lambert HIV-1 infection Cidofovir Gilead Science CMV retinitis, herpes, papillomavirus Curdlan sulfate AJI Pharma USA HIV infection Cytomegalovirus MedImmune CMV retinitis Immune globin Cytovene Syntex Sight threatening Ganciclovir CMV peripheral CMV retinitis Darunavir Tibotec- J & J HIV infection, AIDS, ARC (protease inhibitor) Delaviridine Pharmacia-Upjohn HIV infection, AIDS, ARC (RT inhibitor) Dextran Sulfate Ueno Fine Chem. AIDS, ARC, HIV Ind. Ltd. (Osaka, positive Japan) asymptomatic ddC Hoffman-La Roche HIV infection, AIDS, Dideoxycytidine ARC ddI Bristol-Myers Squibb HIV infection, AIDS, Dideoxyinosine ARC; combination with AZT/d4T DMP-450 AVID HIV infection, (Camden, NJ) AIDS, ARC (protease inhibitor) Efavirenz Bristol Myers Squibb HIV infection, (DMP 266, SUSTIVA ®) AIDS, ARC (−)6-Chloro-4-(S)- (non-nucleoside RT cyclopropylethynyl- inhibitor) 4(S)-trifluoro- methyl-1,4-dihydro- 2H-3,1-benzoxazin- 2-one, STOCRINE EL10 Elan Corp, PLC HIV infection (Gainesville, GA) Etravirine Tibotec/J & J HIV infection, AIDS, ARC (non-nucleoside reverse transcriptase inhibitor) Famciclovir Smith Kline herpes zoster, herpes simplex GS 840 Gilead HIV infection, AIDS, ARC (reverse transcriptase inhibitor) HBY097 Hoechst Marion HIV infection, Roussel AIDS, ARC (non-nucleoside reverse transcriptase inhibitor) Hypericin VIMRx Pharm. HIV infection, AIDS, ARC Recombinant Human Triton Biosciences AIDS, Kaposi's Interferon Beta (Almeda, CA) sarcoma, ARC Interferon alfa-n3 Interferon Sciences ARC, AIDS Indinavir Merck HIV infection, AIDS, ARC, asymptomatic HIV positive, also in combination with AZT/ddI/ddC ISIS 2922 ISIS Pharmaceuticals CMV retinitis KNI-272 Nat'l Cancer Institute HIV-assoc. diseases Lamivudine, 3TC Glaxo Wellcome HIV infection, AIDS, ARC (reverse transcriptase inhibitor); also with AZT Lobucavir Bristol-Myers Squibb CMV infection Nelfinavir Agouron HIV infection, Pharmaceuticals AIDS, ARC (protease inhibitor) Nevirapine Boeheringer Ingelheim HIV infection, AIDS, ARC (RT inhibitor) Novapren Novaferon Labs, Inc. HIV inhibitor (Akron, OH) Peptide T Peninsula Labs AIDS Octapeptide (Belmont, CA) Sequence Trisodium Astra Pharm. CMV retinitis, HIV Phosphonoformate Products, Inc. infection, other CMV infections PNU-140690 Pharmacia Upjohn HIV infection, AIDS, ARC (protease inhibitor) Probucol Vyrex HIV infection, AIDS RBC-CD4 Sheffield Med. HIV infection, Tech (Houston, TX) AIDS, ARC Ritonavir Abbott HIV infection, AIDS, ARC (protease inhibitor) Saquinavir Hoffmann- HIV infection, LaRoche AIDS, ARC (protease inhibitor) Stavudine; d4T Bristol-Myers Squibb HIV infection, AIDS, Didehydrodeoxy- ARC Thymidine Tipranavir Boehringer Ingelheim HIV infection, AIDS, ARC (protease inhibitor) Valaciclovir Glaxo Wellcome Genital HSV & CMV infections Virazole Viratek/ICN asymptomatic HIV Ribavirin (Costa Mesa, CA) positive, LAS, ARC VX-478 Vertex HIV infection, AIDS, ARC Zalcitabine Hoffmann-LaRoche HIV infection, AIDS, ARC, with AZT Zidovudine; AZT Glaxo Wellcome HIV infection, AIDS, ARC, Kaposi's sarcoma, in combination with other therapies Tenofovir disoproxil, Gilead HIV infection, fumarate salt (VIREAD ®) AIDS, (reverse transcriptase inhibitor) EMTRIVA ® Gilead HIV infection, (Emtricitabine) (FTC) AIDS, (reverse transcriptase inhibitor) COMBIVIR ® GSK HIV infection, AIDS, (reverse transcriptase inhibitor) Abacavir succinate GSK HIV infection, (or ZIAGEN ®) AIDS, (reverse transcriptase inhibitor) REYATAZ ® Bristol-Myers Squibb HIV infection (or atazanavir) AIDs, protease inhibitor FUZEON ® Roche/Trimeris HIV infection (Enfuvirtide or T-20) AIDs, viral Fusion inhibitor LEXIVA ® GSK/Vertex HIV infection (or Fosamprenavir calcium) AIDs, viral protease inhibitor Selzentry Pfizer HIV infection AIDs, Maraviroc; (UK 427857) (CCR5 antagonist, in development) (CCR5 antagonist, in development) TRIZIVIR ® GSK HIV infection AIDs, (three drug combination) Sch-417690 (vicriviroc) Schering-Plough HIV infection AIDs, (CCR5 antagonist, in development) TAK-652 Takeda HIV infection AIDs, (CCR5 antagonist, in development) GSK 873140 GSK/ONO HIV infection (ONO-4128) AIDs, (CCR5 antagonist, in development) Integrase Inhibitor Merck HIV infection MK-0518 AIDs Raltegravir TRUVADA ® Gilead Combination of Tenofovir disoproxil fumarate salt (VIREAD ®) and EMTRIVA ® (Emtricitabine) Integrase Inhibitor Gilead/Japan Tobacco HIV Infection GS917/JTK-303 AIDs Elvitegravir in development Triple drug combination Gilead/Bristol-Myers Squibb Combination of Tenofovir ATRIPLA ® disoproxil fumarate salt (VIREAD ®), EMTRIVA ® (Emtricitabine), and SUSTIVA ® (Efavirenz) FESTINAVIR ® Oncolys BioPharma HIV infection 4′-ethynyl-d4T BMS AIDs in development CMX-157 Chimerix HIV infection Lipid conjugate of AIDs nucleotide tenofovir GSK1349572 GSK HIV infection Integrase inhibitor AIDs

Immunomodulators

Drug Name Manufacturer Indication AS-101 Wyeth-Ayerst AIDS Bropirimine Pharmacia Upjohn Advanced AIDS Acemannan Carrington Labs, Inc. AIDS, ARC (Irving, TX) CL246,738 Wyeth AIDS, Kaposi's Lederle Labs sarcoma FP-21399 Fuki ImmunoPharm Blocks HIV fusion with CD4+ cells Gamma Interferon Genentech ARC, in combination w/TNF (tumor necrosis factor) Granulocyte Genetics Institute AIDS Macrophage Colony Sandoz Stimulating Factor Granulocyte Hoechst-Roussel AIDS Macrophage Colony Immunex Stimulating Factor Granulocyte Schering-Plough AIDS, Macrophage Colony combination Stimulating Factor w/AZT HIV Core Particle Rorer Seropositive HIV Immunostimulant IL-2 Cetus AIDS, in combination Interleukin-2 w/AZT IL-2 Hoffman-LaRoche AIDS, ARC, HIV, in Interleukin-2 Immunex combination w/AZT IL-2 Chiron AIDS, increase in Interleukin-2 CD4 cell counts (aldeslukin) Immune Globulin Cutter Biological Pediatric AIDS, in Intravenous (Berkeley, CA) combination w/AZT (human) IMREG-1 Imreg AIDS, Kaposi's (New Orleans, LA) sarcoma, ARC, PGL IMREG-2 Imreg AIDS, Kaposi's (New Orleans, LA) sarcoma, ARC, PGL Imuthiol Diethyl Merieux Institute AIDS, ARC Dithio Carbamate Alpha-2 Schering Plough Kaposi's sarcoma Interferon w/AZT, AIDS Methionine- TNI Pharmaceutical AIDS, ARC Enkephalin (Chicago, IL) MTP-PE Ciba-Geigy Corp. Kaposi's sarcoma Muramyl-Tripeptide Granulocyte Amgen AIDS, in combination Colony Stimulating w/AZT Factor Remune Immune Response Immunotherapeutic Corp. rCD4 Genentech AIDS, ARC Recombinant Soluble Human CD4 rCD4-IgG AIDS, ARC hybrids Recombinant Biogen AIDS, ARC Soluble Human CD4 Interferon Hoffman-La Roche Kaposi's sarcoma Alfa 2a AIDS, ARC, in combination w/AZT SK&F106528 Smith Kline HIV infection Soluble T4 Thymopentin Immunobiology HIV infection Research Institute (Annandale, NJ) Tumor Necrosis Genentech ARC, in combination Factor; TNF w/gamma Interferon

Anti-Infectives

Drug Name Manufacturer Indication Clindamycin with Pharmacia Upjohn Pneumocystic Pneumonia Primaquine PCP Fluconazole Pfizer Cryptococcal meningitis, candidiasis Pastille Squibb Corp. Prevention of Nystatin Pastille oral candidiasis Ornidyl Merrell Dow PCP Eflornithine Pentamidine LyphoMed PCP treatment Isethionate (IM & IV) (Rosemont, IL) Trimethoprim Antibacterial Trimethoprim/sulfa Antibacterial Piritrexim Burroughs Wellcome PCP treatment Pentamidine Fisons Corporation PCP prophylaxis Isethionate for Inhalation Spiramycin Rhone-Poulenc Cryptosporidial diarrhea Intraconazole- Janssen-Pharm. Histoplasmosis; R51211 cryptococcal meningitis Trimetrexate Warner-Lambert PCP Daunorubicin NeXstar, Sequus Kaposi's sarcoma Recombinant Human Ortho Pharm. Corp. Severe anemia Erythropoietin assoc. with AZT therapy Recombinant Human Serono AIDS-related Growth Hormone wasting, cachexia Megestrol Acetate Bristol-Myers Squibb Treatment of anorexia assoc. W/AIDS Testosterone Alza, Smith Kline AIDS-related wasting Total Enteral Norwich Eaton Diarrhea and Nutrition Pharmaceuticals malabsorption related to AIDS

Additionally, the compounds herein set forth can be used in combination with HIV entry inhibitors. Examples of such HIV entry inhibitors are discussed in DRUGS OF THE FUTURE 1999, 24(12), pp. 1355-1362; CELL, Vol. 9, pp. 243-246, Oct. 29, 1999; and DRUG DISCOVERY TODAY, Vol. 5, No. 5, May 2000, pp. 183-194 and Inhibitors of the entry of HIV into host cells, Meanwell, Nicholas A.; Kadow, John F. Current Opinion in Drug Discovery & Development (2003), 6(4), 451-461. Specifically, the compounds can be utilized in combination with attachment inhibitors, fusion inhibitors, and chemokine receptor antagonists aimed at either the CCR5 or CXCR4 co-receptor. HIV attachment inhibitors are also set forth in U.S. Pat. No. 7,354,924 and US 2005/0209246.

It will be understood that the scope of combinations of the compounds of this application with AIDS antivirals, immunomodulators, anti-infectives, HIV entry inhibitors or vaccines is not limited to the list in the above Table but includes, in principle, any combination with any pharmaceutical composition useful for the treatment of AIDS.

Preferred combinations are simultaneous or alternating treatments with a compound of the present disclosure and an inhibitor of HIV protease and/or a non-nucleoside inhibitor of HIV reverse transcriptase. An optional fourth component in the combination is a nucleoside inhibitor of HIV reverse transcriptase, such as AZT, 3TC, ddC or ddI. A preferred inhibitor of HIV protease is REYATAZ® (active ingredient Atazanavir). Typically, a dose of 300 to 600 mg is administered once a day. This can be co-administered with a low dose of Ritonavir (50 to 500 mgs). Another preferred inhibitor of HIV protease is KALETRA®. Another useful inhibitor of HIV protease is indinavir, which is the sulfate salt of N-(2(R)-hydroxy-1-(S)-indanyl)-2(R)-phenylmethyl-4-(S)-hydroxy-5-(1-(4-(3-pyridyl-methyl)-2(S)—N′-(t-butylcarboxamido)-piperazinyl))-pentaneamide ethanolate, and is synthesized according to U.S. Pat. No. 5,413,999. Indinavir is generally administered at a dosage of 800 mg three times a day. Other preferred protease inhibitors are nelfinavir and ritonavir. Another preferred inhibitor of HIV protease is saquinavir which is administered in a dosage of 600 or 1200 mg tid. Preferred non-nucleoside inhibitors of HIV reverse transcriptase include efavirenz. These combinations may have unexpected effects on limiting the spread and degree of infection of HIV. Preferred combinations include those with the following (1) indinavir with efavirenz, and, optionally, AZT and/or 3TC and/or ddI and/or ddC; (2) indinavir, and any of AZT and/or ddI and/or ddC and/or 3TC, in particular, indinavir and AZT and 3TC; (3) stavudine and 3TC and/or zidovudine; (4) tenofovir disoproxil fumarate salt and emtricitabine.

In such combinations the compound of the present invention and other active agents can be administered separately or in conjunction. In addition, the administration of one element can be prior to, concurrent to, or subsequent to the administration of other agent(s).

In another embodiment, these methods are useful for inhibiting viral replication in a patient. Such methods can be useful in treating or preventing HIV disease.

The crystalline forms of the disclosure can also be used as laboratory reagents. The crystalline forms can be instrumental in providing research tools for designing of viral replication assays, validation of animal assay systems and structural biology studies to further enhance knowledge of the HIV disease mechanisms.

The crystalline forms of this disclosure can also be used to treat or prevent viral contamination of materials and therefore reduce the risk of viral infection of laboratory or medical personnel or patients who come in contact with such materials, e.g., blood, tissue, surgical instruments and garments, laboratory instruments and garments, and blood collection or transfusion apparatuses and materials.

Examples

The following non-limiting examples are illustrative of the disclosure.

Preparation of Form H-4 of Compound I

To a 1 L ChemGlass reactor was added tetrahydrofuran (200 mL, 4 L/Kg), Compound I (50 g, 70.9 mmol), and tetrabutylammonium hydroxide (55 wt % in water, 52 mL, 110 mmol) and stirred at 20° C. for 1 hour. After sampling and confirmation of reaction completion by HPLC, hydrochloric acid (6 N in water, 38.5 mL, 213 mmol) was added, followed by acetonitrile (200 mL, 4 L/Kg) and the solution was heated to 40° C. Water (195 mL, 3.9 L/Kg) was added while maintaining the temperature above 35° C. to bring the solvent system to a 4:5:4 volume:volume:volume ratio of THF:water; acetonitrile. Seed crystals of form H-4 of Compound I (2.5 g, 3.35 mmol, jet-milled) were added. The resultant slurry was then wet-milled until reaching a desaturation endpoint as confirmed by HPLC quantitative analysis. Wet-milling was halted and water (350 mL, 7 L/Kg) was added over 4 hours. The slurry was then cooled to 20° C. over a period of 1 hour. The slurry was then filtered and washed with acetonitrile:water (1:3, 200 mL, 4 L/Kg). The wet filter cake was then dried in a vacuum oven at 50° C. and 100-150 torr, while nitrogen with a relative humidity between 8 and 10% was supplied to the oven until drying was complete as confirmed by GC, KF, and PXRD. The H-4 form of Compound I was isolated as a white solid.

Form H-4 was analyzed and characterized using one or more of the methods described herein.

Preparation of Form TIF-1 of Compound I

To a 250 mL ChemGlass reactor was added Compound I (3.0 g, 4.1 mmol), ethanol (63 mL, 21 L/Kg) tetrabutylammonium hydroxide (55 wt % in water, 3.0 mL, 6.2 mmol), and water (15 mL, 5 L/Kg) and the reaction was heated to 63° C. and stirred for 17 hours. After sampling and confirmation of reaction completion by HPLC the reaction mixture was cooled to 20° C. and hydrochloric acid (6 N in water, 2.1 mL, 12 mmol) was added. The slurry was heated to 78° C. then cooled to 50° C. over 2 hours. Water (15 mL, 5 L/Kg) was added over 1 hour and the slurry was maintained at 50° C. for an additional period of 1 hour. The slurry was then cooled to 0° C. over 5 hours. The slurry was then filtered and washed with ethanol:water (3:2, 9 mL, 3 L/Kg). The wet filter cake was dried in a vacuum oven at 50° C. and 100 torr until drying was complete as confirmed by PXRD. Form T1F-1 of Compound I (T1F-1) was isolated as a white solid.

Form T1F-1 was analyzed and characterized using one or more of the methods described herein.

Preparation of Form TIF-2 of Compound I

To a 1 L ChemGlass reactor was added Compound I (40 g, 56.5 mmol), 2-propanol (400 mL, 10 L/Kg), tetrabutylammonium hydroxide (55 wt % in water, 41 mL, 84 mmol) and water (160 mL, 4 L/Kg) and the mixture was heated to 60° C. for 16 hours. After sampling and confirmation of reaction completion by HPLC the reaction mixture was cooled to 20° C., then hydrochloric acid (6 N in water, 30.5 mL, 183 mmol) and 2-propanol (200 mL, 5 L/Kg) were added. The slurry was heated to 80° C. where all solids dissolved. The solution was cooled to 70° C. and seed crystals of form T1F-2 of Compound I (400 mg, 0.55 mmol) were added. The resultant slurry was then cooled to 0° C. over 10 hours. The slurry was then filtered and washed with 2-propanol:water (3:1, 120 mL, 3 L/Kg) then 2-propanol (120 mL, 3 L/Kg). The wet filter cake was then dried in a vacuum oven at 60° C. at 100-150 torr until drying is complete as confirmed by loss on drying (LOD) from the thermogravimetric analysis and PXRD. The T1F-2 form of Compound I (T1F-2) was isolated as a white solid.

Form T1F-2 was analyzed and characterized using one or more of the methods described herein.

Single Crystal X-Ray Measurements

For each of the crystal forms, H-4, T1F-1, and T1F-2 of Compound I, a Bruker X8 APEX2 CCD diffractometer equipped with a micro-focusing X-ray generator of Cu Kα radiation, (λ=1.54178 Å) was used to collect diffraction data at room temperature. Indexing and processing of the measured intensity data were carried out with the APEX2 software package/program suite (APEX2 Data collection and processing user interface: APEX2 User Manual, v 1.27; BRUKER AXS, INc., 5465 East Cheryl Parkway, Madison, Wis. 53711 USA). The final unit cell parameters were determined using the entire data set.

The structures were solved by direct methods and refined by the full-matrix least-squares techniques, using the SHELXTL software package (Sheldrick, G. M. SHELXTL. Structure Determination Programs. Version 6.14, Bruker AXS, Madison, Wis., USA.). The function minimized in the refinements was Σw(|F_(o)|−|F_(c)|)². R is defined as Σ∥F_(o)|−|F_(c)∥/Σ|F| while R_(w)=[Σ_(w)(|F_(o)|−|F_(c)|)²/Σ_(w)|Fo|²]^(1/2), where w is an appropriate weighting function based on errors in the observed intensities. Difference Fourier maps were examined at all stages of refinement. All non-hydrogen atoms were refined with anisotropic thermal displacement parameters. The hydrogen atoms associated with hydrogen bonding were located in the final difference Fourier maps while the positions of the other hydrogen atoms were calculated from an idealized geometry with standard bond lengths and angles. All hydrogen atoms were assigned isotropic temperature factors and included in structure factor calculations with fixed parameters.

The crystal data of the H-4, T1F-1, and T1F-2 forms of Compound I are shown in Tables 1, 2 and 3, respectively. The fractional atomic coordinates of the H-4, T1F-1, and T1F-2 forms of Compound I are listed in Tables 4, 5 and 6, respectively. For the unit cell parameters and atomic coordinates, a number shown in round brackets [e.g., 1.234(5)] indicates the precision in the last digit, and is referred to as an estimated standard deviation (esd) or as a standard uncertainty (su). See, P. G. Jones, Crystal structure determination: a critical view, Chem. Soc. Rev., 1984, 13, 157-172, which is incorporated by reference. It should be understood by one of ordinary skill in the art that slight variations in the coordinates are possible and are considered to be within the scope the present disclosure.

TABLE 1 Single Crystal X-Ray Data for Form H-4 Temperature room temperature Wavelength 1.54178 {acute over (Å)} Crystal system, space group Monoclinic, P2₁ Unit cell dimensions a = 8.9877(3) {acute over (Å)} alpha = 90° b = 12.7723(4) {acute over (Å)} beta = 96.875(2)° c = 17.9546(5) {acute over (Å)} gamma = 90° Volume 2046.2(1) {acute over (Å)}³ Z, Calculated density 2, 1.210 g/cm³ Molecules per unit cell 2

TABLE 2 Single Crystal X-Ray Data for Form T1F-1 Temperature room temperature Wavelength 1.54178 {acute over (Å)} Crystal system, space group Orthorhombic, P2₁2₁2 Unit cell dimensions a =17.3307(6) {acute over (Å)} alpha = 90° b = 30.7745(9) {acute over (Å)} beta = 90° c = 7.8210(2) {acute over (Å)} gamma = 90° Volume 4171.3(2) {acute over (Å)}³ Z, Calculated density 4, 1.158 g/cm³ Molecules per unit cell 4

TABLE 3 Single Crystal X-Ray Data for Form T1F-2 Temperature room temperature Wavelength 1.54178 {acute over (Å)} Crystal system, space group Monoclinic, C2 Unit cell dimensions a = 43.967(1) {acute over (Å)} alpha = 90° b = 7.7218(3) {acute over (Å)} beta = 99.045(2)° c = 12.0227(4) {acute over (Å)} gamma = 90° Volume 4031.3(2) {acute over (Å)}³ Z, Calculated density 4, 1.199 g/cm³ Molecules per unit cell 4

TABLE 4 Atomic Coordinates of Form H-4. Atom X Y Z O1 1.5004 0.8359 0.7458 O2 1.3611 0.8378 0.8375 O3 0.7490 0.5188 −0.0043 O4 0.6115 0.4077 −0.1006 O5 0.0642 0.8319 −0.0268 S1 0.6068 0.4882 −0.0451 N1 0.1223 0.6959 0.0964 N2 0.3779 0.6285 0.0294 C1 1.3732 0.8353 0.7720 C2 1.2396 0.8280 0.7148 C3 1.0993 0.8296 0.7386 C4 0.9718 0.8203 0.6871 C5 0.9829 0.8090 0.6114 C6 1.1235 0.8108 0.5889 C7 1.2515 0.8205 0.6394 C8 0.8450 0.7985 0.5554 C9 0.7806 0.8852 0.5285 C10 0.6486 0.8891 0.4680 C11 0.6397 0.7896 0.4191 C12 0.6472 0.6957 0.4729 C13 0.7877 0.6904 0.5333 C14 0.7357 0.6373 0.6035 C15 0.9135 0.6222 0.5081 C16 0.7727 0.7942 0.3731 C17 0.4834 0.7896 0.3702 C18 0.4353 0.6860 0.3281 C19 0.4607 0.5939 0.3843 C20 0.6157 0.5938 0.4287 C21 0.5293 0.6633 0.2640 C22 0.4559 0.8837 0.3177 C23 0.2924 0.8928 0.2861 C24 0.2372 0.7935 0.2449 C25 0.2620 0.6943 0.2963 C26 0.1647 0.7014 0.3614 C27 0.2100 0.5968 0.2496 C28 0.0499 0.6043 0.2096 C29 0.0272 0.7018 0.1617 C30 0.0729 0.7989 0.2082 C31 0.0150 0.8913 0.1565 C32 −0.1349 0.8458 0.1169 C33 −0.1326 0.7265 0.1281 C34 −0.0056 0.9972 0.1917 C35 0.0526 1.0817 0.1642 C36 −0.0938 1.0049 0.2561 C37 0.1168 0.5988 0.0501 C38 0.2268 0.6067 −0.0063 C39 0.4729 0.6736 −0.0220 C40 0.5169 0.5996 −0.0834 C41 0.4893 0.4509 0.0197 C42 0.4531 0.5401 0.0693 Cl1 1.7705 0.8369 0.8650 H5A −0.0230 0.8290 −0.0601 H5B 0.1080 0.8970 −0.0310 H1 1.5682 0.8401 0.7805 H1A 0.0954 0.7498 0.0654 H1B 0.2184 0.7065 0.1153 H3 1.0900 0.8370 0.7894 H4 0.8778 0.8217 0.7038 H6 1.1330 0.8053 0.5380 H7 1.3453 0.8220 0.6224 H9 0.8188 0.9484 0.5481 H10A 0.6578 0.9500 0.4366 H10B 0.5568 0.8963 0.4910 H12 0.5617 0.7045 0.5015 H14A 0.6591 0.6792 0.6218 H14B 0.8193 0.6310 0.6419 H14C 0.6965 0.5690 0.5903 H15A 0.9960 0.6198 0.5473 H15B 0.9464 0.6516 0.4637 H15C 0.8765 0.5526 0.4976 H16A 0.8588 0.8216 0.4037 H16B 0.7481 0.8388 0.3304 H16C 0.7945 0.7250 0.3565 H17 0.4110 0.7983 0.4064 H19A 0.3860 0.5975 0.4190 H19B 0.4462 0.5285 0.3570 H20A 0.6906 0.5848 0.3946 H20B 0.6238 0.5352 0.4633 H21A 0.6334 0.6598 0.2836 H21B 0.5146 0.7181 0.2273 H21C 0.4987 0.5976 0.2409 H22A 0.4867 0.9472 0.3450 H22B 0.5165 0.8767 0.2767 H23A 0.2797 0.9517 0.2519 H23B 0.2327 0.9059 0.3267 H24 0.2992 0.7839 0.2041 H26A 0.0658 0.7246 0.3423 H26B 0.2089 0.7503 0.3982 H26C 0.1588 0.6337 0.3841 H27A 0.2780 0.5855 0.2123 H27B 0.2166 0.5362 0.2824 H28A 0.0288 0.5430 0.1782 H28B −0.0201 0.6049 0.2468 H30 0.0112 0.7988 0.2497 H31 0.0838 0.9000 0.1185 H32A −0.2191 0.8761 0.1384 H32B −0.1453 0.8624 0.0638 H33A −0.2037 0.7056 0.1619 H33B −0.1572 0.6907 0.0805 H35A 0.0383 1.1469 0.1853 H35B 0.1077 1.0760 0.1237 H36A −0.1115 1.0772 0.2666 H36B −0.0391 0.9728 0.2994 H36C −0.1879 0.9696 0.2441 H37A 0.0165 0.5894 0.0243 H37B 0.1407 0.5386 0.0822 H38A 0.2271 0.5414 −0.0339 H38B 0.1957 0.6620 −0.0418 H39A 0.5640 0.6988 0.0069 H39B 0.4218 0.7337 −0.0461 H40A 0.5828 0.6363 −0.1137 H40B 0.4276 0.5793 −0.1160 H41A 0.3969 0.4237 −0.0066 H41B 0.5365 0.3951 0.0507 H42A 0.3899 0.5139 0.1054 H42B 0.5455 0.5648 0.0972

TABLE 5 Atomic Coordinates of Form T1F-1 Atom X Y Z O1 0.4373 0.7332 0.7291 O2 0.5505 0.7385 0.5905 O3 −0.0883 0.4855 −0.4130 O4 −0.1934 0.4292 −0.4560 S1 −0.1156 0.4404 −0.4219 N1 0.1311 0.3184 −0.2541 N2 0.0159 0.3763 −0.3326 C1 0.4912 0.7204 0.6183 C2 0.4635 0.6806 0.5234 C3 0.3938 0.6608 0.5577 C4 0.3745 0.6244 0.4670 C5 0.4205 0.6062 0.3419 C6 0.4906 0.6273 0.3094 C7 0.5115 0.6641 0.3980 C8 0.3981 0.5658 0.2532 C9 0.4073 0.5289 0.3376 C10 0.3841 0.4842 0.2802 C11 0.3207 0.4863 0.1453 C12 0.3469 0.5202 0.0085 C13 0.3650 0.5668 0.0755 C14 0.4258 0.5865 −0.0462 C15 0.2930 0.5959 0.0735 C16 0.2455 0.4997 0.2363 C17 0.3169 0.4414 0.0569 C18 0.2700 0.4374 −0.1169 C19 0.2962 0.4744 −0.2342 C20 0.2952 0.5185 −0.1513 C21 0.1828 0.4441 −0.0812 C22 0.2959 0.4035 0.1784 C23 0.3071 0.3598 0.1005 C24 0.2602 0.3554 −0.0667 C25 0.2849 0.3912 −0.1985 C26 0.3717 0.3848 −0.2466 C27 0.2376 0.3856 −0.3635 C28 0.2368 0.3398 −0.4383 C29 0.2130 0.3060 −0.3030 C30 0.2620 0.3098 −0.1469 C31 0.2429 0.2702 −0.0367 C32 0.2142 0.2377 −0.1716 C33 0.2170 0.2598 −0.3473 C34 0.3054 0.2517 0.0731 C35 0.2869 0.2356 0.2265 C36 0.3847 0.2548 0.0290 C37 0.0675 0.3034 −0.3675 C38 −0.0026 0.3301 −0.3257 C39 −0.0292 0.4046 −0.2062 C40 −0.0918 0.4200 −0.2330 C41 −0.0599 0.4076 −0.5590 C42 0.0134 0.3962 −0.4970 Cl1 0.4391 0.8231 0.8772 H1 0.4524 0.7547 0.7807 H1A 0.1219 0.3080 −0.1484 H1B 0.1287 0.3475 −0.2478 H3 0.3607 0.6719 0.6405 H4 0.3275 0.6111 0.4910 H6 0.5236 0.6163 0.2262 H7A 0.5580 0.6778 0.3737 H9 0.4307 0.5308 0.4443 H10A 0.3658 0.4676 0.3778 H10B 0.4286 0.4693 0.2333 H12 0.3968 0.5092 −0.314 H14A 0.4337 0.6165 −0.0180 H14B 0.4079 0.5842 −0.1620 H14C 0.4735 0.5709 −0.0343 H15A 0.3074 0.6252 0.1013 H15B 0.2565 0.5855 0.1563 H15C 0.2700 0.5953 −0.0381 H16A 0.2281 0.4764 0.3080 H16B 0.2066 0.5062 0.1526 H16C 0.2548 0.5250 0.3051 H17 0.3704 0.4355 0.0239 H19A 0.3482 0.4683 −0.2735 H19B 0.2628 0.4751 −0.3338 H20A 0.2427 0.5257 −0.1193 H20B 0.3128 0.5400 −0.2330 H21A 0.1551 0.4445 −0.1875 H21B 0.1754 0.4713 −0.0230 H21C 0.1640 0.4208 −0.0112 H22A 0.3274 0.4055 0.2805 H22B 0.2424 0.4064 0.2127 H23A 0.2907 0.3376 0.1808 H23B 0.3614 0.3553 0.0764 H24 0.2062 0.3614 −0.0371 H26A 0.3886 0.4088 −0.3158 H26B 0.3776 0.3582 −0.3093 H26C 0.4022 0.3836 −0.1442 H27A 0.1848 0.3943 −0.3404 H27B 0.2579 0.4053 −0.4493 H28A 0.2878 0.3327 −0.4813 H28B 0.2010 0.3387 −0.5335 H30 0.3152 0.3055 −0.1858 H31 0.1992 0.2778 0.0373 H32A 0.2468 0.2121 −0.1716 H32B 0.1618 0.2289 −0.1456 H33A 0.2646 0.2530 −0.4070 H33B 0.1736 0.2511 −0.4177 H35A 0.3256 0.2267 0.3013 H35B 0.2354 0.2332 0.2584 H36A 0.4144 0.2609 0.1298 H36B 0.3917 0.2778 −0.0526 H36C 0.4016 0.2278 −0.0198 H37A 0.0570 0.2729 −0.3483 H37B 0.0816 0.3074 −0.4865 H38A −0.0434 0.3237 −0.4066 H38B −0.0209 0.3227 −0.2121 H39A −0.0340 0.3876 −0.1024 H39B 0.0044 0.4289 −0.1785 H40A −0.992 0.4430 −0.1493 H40B −0.1294 0.3976 −0.2064 H41A −0.0885 0.3811 −0.5829 H41B −0.0534 0.4228 −0.6671 H42A 0.0448 0.4222 −0.4933 H42B 0.0371 0.3765 −0.5782

TABLE 6 Atomic Coordinates of Form T1F-2 Atom X Y Z O1 0.6155 1.0820 0.1009 O2 0.5822 1.2455 −0.0060 O3 0.2787 0.1168 −0.0382 O4 0.3337 0.1749 −0.0050 S1 0.3050 0.1686 0.0375 N1 0.3049 0.3186 0.4890 N2 0.2817 0.2469 0.2620 C1 0.5872 1.1392 0.0659 C2 0.5637 1.0606 0.1252 C3 0.5701 0.9237 0.1974 C4 0.5471 0.8490 0.2465 C5 0.5168 0.9077 0.2238 C6 0.5111 1.0494 0.1545 C7 0.5340 1.1252 0.1056 C8 0.4914 0.8266 0.2758 C9 0.4809 0.9107 0.3572 C10 0.4551 0.8525 0.4166 C11 0.4336 0.7203 0.3479 C12 0.4546 0.5853 0.3029 C13 0.4782 0.6536 0.2291 C14 0.5041 0.5202 0.2388 C15 0.4643 0.6788 0.1036 C16 0.4136 0.8235 0.2542 C17 0.4151 0.6269 0.4317 C18 0.3971 0.4593 0.3883 C19 0.4193 0.3415 0.3357 C20 0.4352 0.4335 0.2500 C21 0.3698 0.5018 0.2964 C22 0.3950 0.7483 0.4892 C23 0.3818 0.6617 0.5841 C24 0.3644 0.4968 0.5448 C25 0.3855 0.3670 0.4916 C26 0.4124 0.3075 0.5815 C27 0.3667 0.2018 0.4521 C28 0.3488 0.1221 0.5399 C29 0.3289 0.2557 0.5863 C30 0.3486 0.4098 0.6338 C31 0.3270 0.5173 0.6794 C32 0.3034 0.3819 0.7276 C33 0.3128 0.2083 0.6855 C34 0.3415 0.6174 0.7994 C35 0.3262 0.7623 0.8278 C36 0.3684 0.5623 0.8688 C37 0.2766 0.2130 0.4593 C38 0.2610 0.2679 0.3455 C39 0.2731 0.3662 0.1662 C40 0.2979 0.3732 0.0931 C41 0.3086 0.0399 0.1586 C42 0.2822 0.0697 0.2224 Cl1 0.2908 0.7084 0.4208 H1 0.6273 1.1245 0.0621 H1A 0.2991 0.4263 0.5056 H1B 0.3140 0.3267 0.4273 H3 0.5901 0.8811 0.2134 H4 0.5519 0.7570 0.2961 H6 0.4913 1.0950 0.1405 H7 0.5295 1.2209 0.0589 H9 0.4903 1.0155 0.3800 H10A 0.4431 0.9527 0.4320 H10B 0.4637 0.8009 0.4882 H12 0.4672 0.5370 0.3702 H14A 0.5190 0.5550 0.1925 H14B 0.4958 0.4094 0.2142 H14C 0.5139 0.5124 0.3158 H15A 0.4800 0.7166 0.0623 H15B 0.4483 0.7643 0.0977 H15C 0.4558 0.5710 0.0731 H16A 0.4263 0.9046 0.2219 H16B 0.3979 0.8850 0.2853 H16C 0.4041 0.7453 0.1969 H17 0.4309 0.5865 0.4928 H19A 0.4347 0.2964 0.3951 H19B 0.4078 0.2440 0.2996 H20A 0.4199 0.4757 0.1890 H20B 0.4484 0.3522 0.2186 H21A 0.3770 0.5630 0.2361 H21B 0.3553 0.5727 0.3276 H21C 0.3599 0.3963 0.2680 H22A 0.4071 0.8476 0.5188 H22B 0.3782 0.7902 0.4337 H23A 0.3680 0.7413 0.6136 H23B 0.3985 0.7413 0.6136 H24 0.3481 0.5312 0.4838 H26A 0.4045 0.2672 0.6469 H26B 0.4262 0.4030 0.6020 H26C 0.4234 0.2153 0.5517 H27A 0.3521 0.2303 0.3853 H27B 0.3807 0.1152 0.4308 H28A 0.3360 0.0286 0.5054 H28B 0.3633 0.0738 0.6012 H30 0.3649 0.3635 0.6909 H31 0.3159 0.5997 0.6439 H32A 0.3037 0.3782 0.8084 H32B 0.2827 0.4118 0.6916 H33A 0.2950 0.1358 0.6615 H33B 0.3268 0.1482 0.7434 H35A 0.3338 0.8235 0.8930 H35B 0.3084 0.7991 0.7818 H36A 0.3755 0.6517 0.9221 H36B 0.3840 0.5379 0.8236 H36C 0.3641 0.4594 0.9084 H37A 0.2629 0.2305 0.5145 H37B 0.2818 0.0910 0.4586 H38A 0.2426 0.1986 0.3237 H38B 0.2548 0.3882 0.3480 H39A 0.2698 0.4813 0.1945 H39B 0.2540 0.3278 0.1217 H40A 0.3168 0.4163 0.1371 H40B 0.2918 0.4535 0.0316 H41A 0.3092 −0.0811 0.1375 H41B 0.3279 0.0671 0.2068 H42A 0.2629 0.0447 0.1736 H42B 0.2840 −0.0085 0.2862

Powder X-Ray Diffraction (PXRD)

Powder X-Ray diffraction (PXRD) data were obtained using a Bruker C2 General Area Detector Diffraction System (GADDS). The radiation was Cu Kα (40 KV, 40 mA). The sample-detector distance was 15 cm. Samples were placed in sealed glass capillaries with diameters of ≤1 mm. The capillary was rotated during data collection. Data were collected for approximately 2≤2θ≤32° with a sample exposure time of at least 1000 seconds. The resulting two-dimensional diffraction arcs were integrated to create a traditional 1-dimensional PXRD pattern with a step size of 0.05 degrees 2θ in the approximate range of 2 to 32 degrees 2θ (two theta).

The results of the PXRD pattern and a simulated pattern calculated from the single crystal data are shown in FIGS. 1, 6, and 9, for the H-4, T1F-1, and T1F-2 forms of Compound I, respectively.

Table 7 lists the characteristic PXRD peaks that describe Forms H-4, T1F-1, and T1F-2 of Compound I.

TABLE 7 Characteristic diffraction peak positions (degrees 2θ ± 0.2) at room temperature, based on a high quality pattern collected with a diffractometer (CuKα) with a spinning capillary with 2θ calibrated with a NIST or other suitable standard. Table 7 Characteristic Diffraction Peak Positions Form H-4 Form T1F-1 Form T1F-2 5.0 7.7 4.1 8.5 11.7 7.4 12.6 13.7 11.9 13.9 14.2 13.0 14.9 15.3 14.0 17.1 16.4 14.5 18.3 17.6 15.5

Differential Scanning Calorimetry (DSC)

TA INSTRUMENT® models Q2000, Q1000 or 2920 were used to generate differential scanning calorimetry (DSC) data. The measurement was made using standard TA Instruments hermetic pans. The measurement was made at a heating rate of 10° C./min, in a nitrogen environment from room temperature to 300° C., with a sample size of about 2-10 mg. The DSC plot was made with the endothermic peaks pointing down.

The DSC results are shown in FIGS. 2, 7, and 12 for the H-4, T1F1, and T1F-2 forms of Compound I, respectively. As seen from FIG. 2, the DSC results for the H-4 form demonstrate a broad endotherm up to about 150° C. As seen from FIG. 7, the DSC results for the T1F-1 form demonstrate a broad endotherm near about 275° C. As seen from FIG. 12, the DSC results for the T1F-2 form demonstrate a broad endotherm near about 275° C.

Thermogravimetric Analysis (TGA)

TA INSTRUMENT® models Q5000, Q500 or 2950 were used to generate thermogravimetric analysis (TGA) data. The measurement was made using standard TA Instruments Platinum pans. The measurement was made at a heating rate of 10° C./min, in a nitrogen environment from room temperature to 300° C., with a sample size about 10-30 mg.

The TGA results are shown in FIGS. 3, 8, and 13 for the H-4, T1F1, and T1F-2 forms of Compound I, respectively. As seen from FIG. 3, the TGA results for the H-4 form demonstrate a 2.5% weight loss up to about 150° C. These results are consistent with a theoretical weight loss of 2.4% for a monohydrate. As seen from FIG. 8, the TGA results for the T1F-1 form demonstrate a negligible weight loss of 0.4% up to about 150° C. These results are consistent with a theoretical weight loss of zero for a non-solvated form. As seen from FIG. 13, the TGA results for the T1F-2 form demonstrate a negligible weight loss of 0.2% up to about 150° C. These results are consistent with a theoretical weight loss of zero for a non-solvated form.

Raman and FTIR Spectroscopy

An IS-50 FTIR/Raman spectrometer was used to collect Raman and FTIR spectrum. The Raman spectrum was collected by using 1064 nm laser with 250 mW. The spectral resolution was 4 cm⁻¹ and scanning number was 64.

Raman spectra are shown in FIGS. 4 and 12 for the H-4 and T1F-2 forms of Compound I, respectively. The Raman spectra are presented as intensity versus Raman shift in cm⁻¹. A listing of the most prominent peaks, reported as Raman shift in cm¹, is provided in Tables 8 and 9 for the H-4 and T1F-2 forms of Compound I, respectively.

The FTIR spectra are shown in FIGS. 5 and 13 for the H-4 and T1F-2 forms of Compound I, respectively. The FTIR spectra are presented as log (1/R) versus wavenumber (cm⁻¹), it being understood that log(1/R)=A (where R is reflectance and A is absorbance). A listing of the most prominent absorption bands, reported in wavenumbers (cm⁻¹), is provided in Tables 10 and 11 for the H-4 and T1F-2 forms of Compound I, respectively.

TABLE 8 Raman Characteristic Peaks for Form H-4 Wavelength (cm⁻¹) 2915.2 1709.3 1660.4 1605.9 676.6

TABLE 9 Raman Characteristic Peaks for Form T1F-2 Wavelength (cm⁻¹) 2930.0 1731.9 1653.6 1609.1 668.0

TABLE 10 IR Characteristic Peaks for Form H-4 Wavelength (cm⁻¹) 3540.3 3271.3 1705.6 1605.5 1229.0 501.8

TABLE 11 IR Characteristic Peaks for Form T1F-2 Wavelength (cm⁻¹) 3372.6 1730.1 1608.6 1575.6 1200.4 494.8

INCORPORATION BY REFERENCE

The entire disclosure of each of the patent documents, including certificates of correction, patent application documents, scientific articles, governmental reports, websites, and other references that may be referred to herein is incorporated by reference herein in its entirety for all purposes. In case of a conflict in terminology, the present specification controls.

EQUIVALENTS

The invention can be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are to be considered in all respects illustrative rather than limiting on the invention described herein. In the various embodiments of the present invention, where the term “comprising” or “comprises” is used, it is also contemplated that in certain embodiments the term “consisting essentially of” or “consists essentially of” can be used, and it is also contemplated that in other certain embodiments the term “consisting of” or “consists of” can be used.

In the specification, the singular forms also include the plural forms, unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In the case of conflict, the present specification will control.

All percentages and ratios used herein, unless otherwise indicated, are by weight. Also, throughout the disclosure the term “weight” is used. It is recognized the mass of an object is often referred to as its weight in everyday usage and for most common scientific purposes, but that mass technically refers to the amount of matter of an object, whereas weight refers to the force experienced by an object due to gravity. Also, in common usage the “weight” (mass) of an object is what one determines when one “weighs” (masses) an object on a scale or balance. 

1. Form H-4, T1F-1, or T1F-2 of Compound I.


2. Form H-4 of Compound I


3. Form H-4 of Compound I according to claim 2 characterized by the following unit cell parameters: Cell Dimensions: a=8.9877(3) Å b=12.7723(4) Å c=17.9546(5) Å α=90 degrees β=96.875(2) degrees γ=90 degrees Space group Monoclinic, P2₁ Molecules/unit cell 2, wherein measurement of said crystalline form is at a temperature between about 20° C. to about 25° C.
 4. Form H-4 of Compound I of claim 2 characterized by fractional atomic coordinates within the unit cell as listed in Table
 4. 5. Form H-4 of Compound I of according to claim 2 with characteristic peak positions in the powder X-Ray diffraction pattern at values of two theta of 5.0±0.2, 8.5±0.2, 12.6±0.2, 13.9±0.2, 14.9±0.2, 17.1±0.2, and 18.3±0.2 at a temperature between about 20° C. and about 25° C.
 6. Form H-4 of Compound I of according to claim 2 characterized by an experimental powder X-ray diffraction pattern substantially in accordance with that shown in FIG.
 1. 7. Form H-4 of Compound I according to claim 2 characterized by a differential scanning calorimetry thermogram substantially in accordance with that shown in FIG.
 2. 8. Form H-4 of Compound I according to claim 7 having a broad endotherm observed up to about 150° C.
 9. Form H-4 of Compound I according to claim 2 characterized by a thermogravimetric analysis plot substantially in accordance with that shown in FIG.
 3. 10. Form H-4 of Compound I according to claim 9 characterized by about a 2.4 percent weight loss up to about 150° C.
 11. A substantially pure form of H-4 of Compound I according to claim
 2. 12. A substantially pure form of H-4 of Compound I according to claim 11 having a purity of at least 95 weight percent.
 13. A substantially pure form of H-4 of Compound I according to claim 11 having a purity of at least 99 weight percent.
 14. A pharmaceutical composition comprising form H-4 of Compound I according to claim 2 and a pharmaceutically acceptable carrier or diluent.
 15. A pharmaceutical composition comprising form H-4 of Compound I according to claim 14 and a pharmaceutically acceptable carrier or diluent.
 16. A pharmaceutical composition comprising form H-4 of Compound I according to claim 12 and a pharmaceutically acceptable carrier or diluent.
 17. A pharmaceutical composition comprising form H-4 of Compound I according to claim 13 and a pharmaceutically acceptable carrier or diluent.
 18. A pharmaceutical composition comprising Form H-4 of Compound I according to claim 2 in combination with one or two additional compounds having anti-HIV activity. 19-67. (canceled) 